Characterization of currently marketed heparin products: analysis of molecular weight and heparinase-I digest patterns

We evaluated polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) approaches to determine weight-average molecular weight (M _w) and polydispersity (PD) of heparins. A set of unfractionated heparin sodium (UFH) and low-molecular-weight heparin (LMWH) samples obtained from nine manufacturers which supply the US market were assessed. For SEC-MALLS, we measured values for water content, refractive index increment (dn/dc), and the second virial coefficient (A ₂) for each sample prior to molecular weight assessment. For UFH, a mean ± standard deviation value for M _w of 16,773 ± 797 was observed with a range of 15,620 to 18,363 (n = 20, run in triplicate). For LMWHs by SEC-MALLS, we measured mean M _w values for dalteparin, tinzaparin, and enoxaparin of 6,717 ± 71 (n = 4), 6,670 ± 417 (n = 3), and 3,959 ± 145 (n = 3), respectively. PAGE analysis of the same UFH, dalteparin, tinzaparin, and enoxaparin samples showed values of 16,135 ± 643 (n = 20), 5,845 ± 45 (n = 4), 6,049 ± 95 (n = 3), and 4,772 ± 69 (n = 3), respectively. These orthogonal measurements are the first M _w results obtained with a large heparin sample set on product being marketed after the heparin crisis of 2008 changed the level of scrutiny of this drug class. In this study, we compare our new data set to samples analyzed over 10 years earlier. In addition, we found that the PAGE analysis of heparinase digested UFH and neat LMWH samples yield characteristic patterns that provide a facile approach for identification and assessment of drug quality and uniformity.     

Resonances and wave propagation velocity in the subglottal airways

Previous studies of subglottal resonances have reported findings based on relatively few subjects, and the relations between these resonances, subglottal anatomy, and models of subglottal acoustics are not well understood. In this study, accelerometer signals of subglottal acoustics recorded during sustained [a:] vowels of 50 adult native speakers (25 males, 25 females) of American English were analyzed. The study confirms that a simple uniform tube model of subglottal airways, closed at the glottis and open at the inferior end, is appropriate for describing subglottal resonances. The main findings of the study are (1) whereas the walls may be considered rigid in the frequency range of Sg2 and Sg3, they are yielding and resonant in the frequency range of Sg1, with a resulting ~4/3 increase in wave propagation velocity and, consequently, in the frequency of Sg1; (2) the "acoustic length" of the equivalent uniform tube varies between 18 and 23.5 cm, and is approximately equal to the height of the speaker divided by an empirically determined scaling factor; (3) trachea length can also be predicted by dividing height by another empirically determined scaling factor; and (4) differences between the subglottal resonances of males and females can be accounted for by height-related differences.     

Superbosonization of Invariant Random Matrix Ensembles

‘Superbosonization’ is a new variant of the method of commuting and anti-commuting variables as used in studying random matrix models of disordered and chaotic quantum systems. We here give a concise mathematical exposition of the key formulas of superbosonization. Conceived by analogy with the bosonization technique for Dirac fermions, the new method differs from the traditional one in that the superbosonization field is dual to the usual Hubbard-Stratonovich field. The present paper addresses invariant random matrix ensembles with symmetry group U _n , O _n , or USp _n , giving precise definitions and conditions of validity in each case. The method is illustrated at the example of Wegner’s n-orbital model. Superbosonization promises to become a powerful tool for investigating the universality of spectral correlation functions for a broad class of random matrix ensembles of non-Gaussian and/or non-invariant type.     

On a quadratic first integral for the charged particle orbits in the charged Kerr solution

Associated with the charged Kerr solution of the Einstein gravitational field equation there is a Killing tensor of valence two. The Killing tensor, which is related to the angular momentum of the field source, is shown to yield a quadratic first integral of the equation of the motion for charged test particles.The research reported herein has been supported in part by the National Science Foundation, grants GP-8868, GP-20023, and GU-1598; the Air Force Office of Scientific Research, grant 903-67; the National Aeronautics and Space Administration, grant 44-004-001; and the Westinghouse Corporation.     

On the orientational dependence of giant magnetoresistance

The functional dependence of the giant magnetoresistance (GMR) with respect to the relative angle between the orientations of the magnetization in the magnetic slabs of a trilayer system is calculated by using the Kubo-Greenwood formula for electrical transport together with the fully-relativistic spin-polarized screened Korringa-Kohn-Rostoker method for semi-infinite systems and the coherent potential approximation. It is found that the functional dependence of the GMR is essentially of the form . Received 30 November 1998     

Methodology for calculating the volume of condensate droplets on topographically modified, microgrooved surfaces

Liquid droplets on micropatterned surfaces consisting of parallel grooves tens of micrometers in width and depth are considered, and a method for calculating the droplet volume on these surfaces is presented. This model, which utilizes the elongated and parallel-sided nature of droplets condensed on these microgrooved surfaces, requires inputs from two droplet images at ? = 0° and ? = 90°--namely, the droplet major axis, minor axis, height, and two contact angles. In this method, a circular cross-sectional area is extruded the length of the droplet where the chord of the extruded circle is fixed by the width of the droplet. The maximum apparent contact angle is assumed to occur along the side of the droplet because of the surface energy barrier to wetting imposed by the grooves--a behavior that was observed experimentally. When applied to water droplets condensed onto a microgrooved aluminum surface, this method was shown to calculate the actual droplet volume to within 10% for 88% of the droplets analyzed. This method is useful for estimating the volume of retained droplets on topographically modified, anisotropic surfaces where both heat and mass transfer occur and the surface microchannels are aligned parallel to gravity to assist in condensate drainage.     

Perpendicular transport in Fe/InP/Fe heterostructures

Perpendicular electric transport in Fe/InP/Fe heterostructures with different terminations is investigated within the relativistic spin-polarized version of the screened Korringa–Kohn–Rostoker method and the Kubo–Greenwood formula, and compared to a Landauer-like approach. Both methods show that the magnetoresistance becomes constant with increasing spacer thickness.     

Analytical techniques and bioactivity assays to compare the structure and function of filgrastim (granulocyte-colony stimulating factor) therapeutics from different manufacturers

The FDA has approved more than 100 protein and peptide drugs with hundreds more in the pipeline (Lanthier et al. in Nat Rev Drug Discov 7(9):733–737, 2008). Many of these originator biologic products are now coming off patent and are being manufactured by alternate methods than the innovator as follow-on drugs. Because changes to the production method often lead to subtle differences (e.g., degradation products, different posttranslational modifications or impurities) in the therapeutic (Schiestl et al. in Nat Biotechnol 29(4):310–312, 2011), there is a critical need to define techniques to test and insure the quality of these drugs. In addition, the emergence of protein therapeutics manufactured by unapproved methodologies presents an ongoing and growing regulatory challenge. In this work, high-resolution mass spectrometry was used to determine the presence or absence of posttranslational modifications for one FDA-approved and three foreign-sourced, unapproved filgrastim products. Circular dichroism (CD) was used to compare the secondary structure and probe the temperature stability of these products. Native 2D ¹H,¹⁵N-heteronuclear singular quantum coherence (HSQC) NMR test was applied to these samples to compare the higher-order structure of the four products. Finally, a cell proliferation assay was performed on the filgrastims to compare their bioactivity, and stressed filgrastim was tested in the bioassay to better understand the effects of changes in protein structure on activity. The results showed that orthogonal approaches are capable of characterizing the physiochemical properties of this protein drug and assessing the impact of structural changes on filgrastim purity and potency.